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OWLS APPLICATION NOTES NO-010

EC-OWLS FOR STUDYING THE EFFECT OF DIFFERENT STRESS FACTORS ON LACTIC ACID BACTERIA
Abstract
Lactic acid bacteria are part of the normal human microflora and play an important role in the fermentation of different food products, such as milk, cheese or meats. During the fermentation process lactobacilli are confronted with many inhibitor factors, such as high salt content, temperature changes, organic acids. It is of great importance to study the effect of different stress factors on the microbial status of lactic acid bacteria, their growth, viability and survival. Lactobacillus plantarum 2142 (Culture Collection of the Institute of Dairy Microbiology, University of P e r u g i a ) wa s u s e d . F r o z e n c u l t u r e o f t h e l a c t o b a c i l l i was transferred in deMan Rogosa Sharpe (MRS) broth (Merck, Darmstadt) using 1% inocula and i n c u b a t e d a t 3 0 °C f o r 2 4 h . B e f o r e e x p e r i m e n t s i t was subcultured at least two times in sterile MRS broth. Electrochemical Optical W aveguide Lightmode Spectroscopy (EC-OW LS) com bines evanescentfield optical sensing with electrochemical control of surface adsorption processes. A layer of indium tin o xi d e ( I T O ) s e r v e s a s a c o n d u c t i v e e l e c t r o d e f o r electrochemical sensing and together with the other glass-type layers of the sensor functioned as a high refractive index waveguide for optical sensing. Mass signal of immobilized cells as function of polarizing potential Effect of acetic acid treatment on the lactobacillus cells Sensor response of different rate of living and damaged cells D i f f e r e n t r a t i o o f l i v i n g a n d h e a t t r e a t e d c e l l s we r e m i xe d a n d m e a s u r e d . U s i n g t h e s a m e o v e r a l l c e l l number, the rate of living cells could be determined.

500 450 Mass (arbitrary unit)….. 400 350 300 250 200 150 100 50 0 50

5 M acetic acid 1M acetic acid 500 mM acetic acid 100 mM acetic acid 20 mM acetic acid 0 acetic acid

60

70

80

90

100

The effect of polarization potential on the number of immobilized cells, measured as the sensor response, was studied. The highest response was o b t a i n e d w h e n 0 . 8 - 1 . 0 V p o t e n t i a l wa s u s e d . A s a result of polarizing potential, the bacterial cells were adsorbed in native form on the surface of the sensor. Calibration curve of lactobacillus cell concentration

Time (min)

Effect of hydrogen lactobacillus cells

peroxide

treatment

on

the

The calibration curve of the bacteria count was obtained from 102 –108 CFU/mL and by using the average of three replicate measurements. According to the calibration curve, a typical first-order kinetic profile in the semi-logarithmic scale can be observed.

References: Adányi, N., Németh E.; Halász A.; Szendrı I.; Váradi M.(2006): Application of electrochemical optical waveguide lightmode spectroscopy (EC-OW LS) for studyi ng the effect of different stress factors on lactic acid bacteria. Anal Chim Acta 573-574. 41-47.

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